Structure prediction and validation of the ERK8 kinase domain.

Abstract

Extracellular signal-regulated kinase 8 (ERK8) has been already implicated in cell transformation and in the protection of genomic integrity and, therefore, proposed as a novel potential therapeutic target for cancer. In the absence of a crystal structure, we developed a three-dimensional model for its kinase domain. To validate our model we applied a structure-based virtual screening protocol consisting of pharmacophore screening and molecular docking. Experimental characterization of the hit compounds confirmed that a high percentage of the identified scaffolds was able to inhibit ERK8. We also confirmed an ATP competitive mechanism of action for the two best-performing molecules. Ultimately, we identified an ERK8 drug-resistant "gatekeeper" mutant that corroborated the predicted molecular binding mode, confirming the reliability of the generated structure. We expect that our model will be a valuable tool for the development of specific ERK8 kinase inhibitors.

• Authors: Angela Strambi, Mattia Mori, Matteo Rossi, David Colecchia, Fabrizio Manetti, Francesca Carlomagno, Maurizio Botta, Mario Chiariello
• Journal: PloS one (PLoS One) Vol. 8 Issue 1 Pg. e52011 ( 2013) ISSN: 1932-6203 [Electronic] United States
• PMID: 23326322 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Validation Study)

Chemical References

• Protein Kinase Inhibitors
• Adenosine Diphosphate
• Adenosine Triphosphate
• Extracellular Signal-Regulated MAP Kinases
• MAPK15 protein, human

Topics

• Adenosine Diphosphate (chemistry, metabolism)
• Adenosine Triphosphate (chemistry, metabolism)
• Amino Acid Sequence
• Binding Sites (genetics)
• Biocatalysis (drug effects)
• Blotting, Western
• Extracellular Signal-Regulated MAP Kinases (chemistry, genetics, metabolism)
• Humans
• Models, Molecular
• Molecular Dynamics Simulation
• Molecular Sequence Data
• Molecular Structure
• Mutation
• Protein Binding
• Protein Kinase Inhibitors (chemistry, metabolism, pharmacology)
• Protein Structure, Tertiary
• Sequence Homology, Amino Acid